Phylogenetic diversity of endolithic bacteria in Bole granite rock in Xinjiang

The endolithic environment is a ubiquitous microbial habitat for microorganisms, such as lichens, Cyanobacteria and fungi, and it provides mineral nutrients and growth surfaces. In extremely environments, such as hot and cold desert, endolithic communities are often the main form of life. More recently, endolithic microbial communities have been observed inhabiting a variety of rock types ranging from hard granite to porous rocks such as basalt, dolomite, limestone, sandstone and granites. Regardless of geographic location and rock type, each of these habitats is characterized by a subsurface microclimate that prevents endolithic microorganisms growth. Photosynthesis-based endolithic microbial communities commonly inhabit the outer millimeters to centimeters of rocks exposed to the surface. The ability to fix carbon dioxide and in some cases atmospheric dinitrogen, gives the Cyanobacteria a clear competitive advantage over heterotrophic bacteria, so it is been called the main primary producer. Light quality and intensity appear to be the main determinant of the maximum depth to which growth occurs in endolithic phototrophic communities. Valleys of Fantastic Rocks in Bole is close to Alashankou Port of Xinjiang which belongs to extreme continental climate. In order to investigate the structure, composition and diversity of endolithic bacterial community in exposed granitic porphyry in the Valleys of Fantastic Rocks, environmental DNA was directly extracted from granite rock, the 16S rRNA genes were amplified from the total DNA by PCR with bacterial-specific primers, and an endolithic bacterial clone library was constructed. Positive clones were randomly selected from the library and identified by Restriction Fragment Length Polymorphism (RFLP). The unique rRNA types clones were sequenced, analysised and then constructed phylogenetic tree. In total, 129 positive clones were screened and grouped into 46 operational taxonomic unites (OTUs). The clone coverage C value was 89.15%, indicating that most of the estimated endolithic bacterial diversity was sampled. BLAST analysis indicated that 46 OTUs were divided into seven phyla (Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, Cyanobacteria, Planctomycetes, Proteobacteria) and five unknown groups. Cyanobacteria (43%), especially the Gp I, form the functional basis for an endolithic bacteria community which contain a wide spectrum species of chemotrophic bacteria (33%) with mainly Actinobacteria, α-Proteobacteria, Acidobacteria. Additionally, most clones that derived from the endolithic bacteria clone library showed high similarity to the sequence deposited in GenBank database with 97%–99%. Besides, 35% of the clones showed less than 97% of sequence similarity, of which 12% sequences were affiliated to genus Rubrobacter. The results suggested that endolithic bacteria in Valleys of Fantastic Rocks in Xinjiang were highly diverse in species richness, and maybe have a diversity of potential novel species and lineages.     

Phylogenetic Composition of Rocky Mountain Endolithic Microbial Ecosystems

The endolithic environment, the pore space in rocks, is a ubiquitous microbial habitat. Photosynthesis-based endolithic communities inhabit the outer few millimeters to centimeters of rocks exposed to the surface. Such endolithic ecosystems have been proposed as simple, tractable models for understanding basic principles in microbial ecology. In order to test previously conceived hypotheses about endolithic ecosystems, we studied selected endolithic communities in the Rocky Mountain region of the United States with culture-independent molecular methods. Community compositions were determined by determining rRNA gene sequence contents, and communities were compared using statistical phylogenetic methods. The results indicate that endolithic ecosystems are seeded from a select, global metacommunity and form true ecological communities that are among the simplest microbial ecosystems known. Statistical analysis showed that biogeographical characteristics that control community composition, such as rock type, are more complex than predicted. Collectively, results of this study support the idea that patterns of microbial diversity found in endolithic communities are governed by principles similar to those observed in macroecological systems.     

Phylogenetic composition of Rocky Mountain endolithic microbial ecosystems

The endolithic environment, the pore space in rocks, is a ubiquitous microbial habitat. Photosynthesis-based endolithic communities inhabit the outer few millimeters to centimeters of rocks exposed to the surface. Such endolithic ecosystems have been proposed as simple, tractable models for understanding basic principles in microbial ecology. In order to test previously conceived hypotheses about endolithic ecosystems, we studied selected endolithic communities in the Rocky Mountain region of the United States with culture-independent molecular methods. Community compositions were determined by determining rRNA gene sequence contents, and communities were compared using statistical phylogenetic methods. The results indicate that endolithic ecosystems are seeded from a select, global metacommunity and form true ecological communities that are among the simplest microbial ecosystems known. Statistical analysis showed that biogeographical characteristics that control community composition, such as rock type, are more complex than predicted. Collectively, results of this study support the idea that patterns of microbial diversity found in endolithic communities are governed by principles similar to those observed in macroecological systems.     

Epilithic and Endolithic Bacterial Communities in Limestone from a Maya Archaeological Site

Biodeterioration of archaeological sites and historic buildings is a major concern for conservators, archaeologists, and scientists involved in preservation of the world's cultural heritage. The Maya archaeological sites in southern Mexico, some of the most important cultural artifacts in the Western Hemisphere, are constructed of limestone. High temperature and humidity have resulted in substantial microbial growth on stone surfaces at many of the sites. Despite the porous natureof limestone and the common occurrence of endolithic microorganisms in many habitats, little is known about the microbial flora living inside the stone. We found a large endolithic bacterial community in limestone from the interior of the Maya archaeological site Ek' Balam. Analysis of 16S rDNA clones demonstrated disparate communities (endolithic: >80% Actinobacteria, Acidobacteria, and Low GC Firmicutes; epilithic: >50% Proteobacteria). The presence of differing epilithic and endolithic bacterial communities may be a significant factor for conservation of stone cultural heritage materials and quantitative prediction of carbonate weathering.     

Geomicrobiology of the ocean crust: a role for chemoautotrophic Fe-bacteria

The delicate balance of the major global biogeochemical cycles greatly depends on the transformation of Earth materials at or near its surface. The formation and degradation of rocks, minerals, and organic matter are pivotal for the balance, maintenance, and future of many of these cycles. Microorganisms also play a crucial role, determining the transformation rates, pathways, and end products of these processes. While most of Earth's crust is oceanic rather than terrestrial, few studies have been conducted on ocean crust transformations, particularly those mediated by endolithic (rock-hosted) microbial communities. The biology and geochemistry of deep-sea and sub-seafloor environments are generally more complicated to study than in terrestrial or near-coastal regimes. As a result, fewer, and more targeted, studies usually homing in on specific sites, are most common. We are studying the role of endolithic microorganisms in weathering seafloor crustal materials, including basaltic glass and sulfide minerals, both in the vicinity of seafloor hydrothermal vents and off-axis at unsedimented (young) ridge flanks. We are using molecular phylogenetic surveys and laboratory culture studies to define the size, diversity, physiology, and distribution of microorganisms in the shallow ocean crust. Our data show that an unexpected diversity of microorganisms directly participate in rock weathering at the seafloor, and imply that endolithic microbial communities contribute to rock, mineral, and carbon transformations.     

Ancient Fungal Life in North Pacific Eocene Oceanic Crust

We present evidence that eukaryotic life has existed in an extreme environment, inside the oceanic crust. Up to now only prokaryotes have been discovered within deep marine sediments and glass-rims of pillow basalts, no higher life forms are described as yet. This study demonstrates unique filamentous fossil structures observed within carbonate-filled vesicles of a massive lava flow unit from the upper oceanic crust in the North Pacific (ODP Site 1224). Based on morphological traits including branching, septa and central pores, the filaments are interpreted as fungi. The chemical composition of the fungal structures differs from the surrounding crystalline carbonate matrix in the deep basaltic rocks. Small open space between the fungi and the carbonate cement and undisturbed filamentous growth through different calcite crystals indicate endolithic fungal growth after the calcium carbonate filling. The presence of euhedral pyrite crystals within the carbonate cements points out anaerobic conditions in this habitat. Our results provide for the first time evidence for eukaryotic, fungal life in deep ocean basaltic rocks.     

Applied microfacies analysis: Provenance studies of Roman mosaic stones

Summary Provenance analysis of archaeological materials is an essential tool of archaeometry but has been rarely applied to antique mosaics. Many mosaics are made of carbonate mosaic stones (tesserae). Hence, microfacies analysis offers a great potential in differentiating these limestone tesserae and provenancing their local, regional, imported or recycled origins. The methods of microfacies analysis and their prospects for studying mosaics are demonstrated by case studies of Roman mosaics from southern Germany (Kraiburg, Bavaria). Austria (Hemmaberg near Globasnitz, southern Carinthia), Italy (Asolo north of Padova) and late Punic and Roman mosaics from Tunisia (Carthage and Hergla). Microfacies-based provenance analysis comprises six stages: 1) Macroscopic assessment and definition of rock colour groups of tesserae, 2) Sampling based on rock-colour categories, 3) Definition of microfacies types and attribution to standard microfacies types based on thin-section criteria, 4) Evaluation of the mosaic site with respect to geological and paleontological data followed by comparisons of the microfacies inventory of the region with microfacies types of the tesserae, 5) Assessment of carbonate tesserae to specific geological rock/time units followed by suggestions of provenance sites. 6) Critical evaluation of provenance assessment within the archaeological context. Principal limestone colours do not necessarily coincide with specific limestone types. Uniformly coloured tesserae can represent different limestone types from different sources (cf. Kraiburg, Hemmaberg). On the other hand, a specific limestone type may be characterized by different rock colours (cf. Carthage). Provenance assessment of mosaic stones results in relatively narrow (Hemmaberg, Asolo) or only broad (Carthage, Hergla) indications of sites characterized by exposures of carbonate rocks whose microfacies criteria, geological age and rock colour correspond to those of the carbonate tesserae. The case studies argue for local and/or imported (Kraiburg) as well as regional provenances of the mosaic material (Hemmaberg, Asolo; source area within a distance between 10 and about 40 km) and indicate that off-cuts of building stones exploited within a regional frame might possibly have been used (Carthage). Differences in the composition and diversity of carbonate rocks used for the fabrication of mosaics reflect time-dependent changes in major quarrying and potential source areas (Carthage). Further provenance research of mosaics should be based on statistically representative tesserae samples, supplemented by microfacies studies of the building material used at the mosaic sites as well as investigation of nonlithic tesserae and the mortar bedding of the mosaics. The isotopic composition of limestone and dolomite tesserae also assists in the critical evaluation of microfacies categorization.     

Nitrogen economy of endolithic microbial communities in hot and cold deserts

The source of combined nitrogen in endolithic microbial communities was studied in samples from desert localities in North and South America, the Middle East, South Africa, and Antarctica. Nitrogen fixation (acetylene reduction) seems to occur only exceptionally. Evidence suggests that, in general, the nitrogen source for endolithic microorganisms in deserts is abiotically fixed nitrogen produced by atmospheric electric discharges (lightning or aurorae), conveyed to the rock by atmospheric precipitation. Nitrogen is apparently not a limiting factor in these low-productivity communities. An incomplete nitrogen cycle seems to be present which includes the following pathways: supply of nitrates and ammonia from the atmosphere; decomposition of organic matter to ammonia; reassimilation of ammonia; ammonia volatilization; loss of organic matter through weathering (only in certain Antarctic rocks); biological nitrogen fixation (exceptional).     

Fungal Diversity Is Not Determined by Mineral and Chemical Differences in Serpentine Substrates

The physico-chemical properties of serpentine soils lead to strong selection of plant species. Whereas many studies have described the serpentine flora, little information is available on the fungal communities dwelling in these sites. Asbestos minerals, often associated with serpentine rocks, can be weathered by serpentine-isolated fungi, suggesting an adaptation to this substrate. In this study, we have investigated whether serpentine substrates characterized by the presence of rocks with distinct mineral composition could select for different fungal communities. Both fungal isolation and 454 pyrosequencing of amplicons obtained from serpentine samples following direct DNA extraction revealed some fungal taxa shared by the four ophiolitic substrates, but also highlighted several substrate-specific taxa. Bootstrap analysis of 454 OTU abundances indicated weak clustering of fungal assemblages from the different substrates, which did not match substrate classification based on exchangeable macronutrients and metals. Intra-substrate variability, as assessed by DGGE profiles, was similar across the four serpentine substrates, and comparable to inter-substrate variability. These findings indicate the absence of a correlation between the substrate (mineral composition and available cations) and the diversity of the fungal community. Comparison of culture-based and culture-independent methods supports the higher taxonomic precision of the former, as complementation of the better performance of the latter.     

Limestone Dissolution Induced by Fungal Mycelia,Acidic Materials,and Carbonic Anhydrase from Fungi

Microorganisms influence the dissolution of a number of minerals. Limestone is one of the most abundant rock types in karst areas, and is predominantly calcium carbonate. Two types of experimental systems were designed in this paper, to make comparisons of limestone dissolution rate among the acidic materials and extracellular carbonic anhydrase (CA) excreted by fungi and the enwrapping effect of fungal mycelia. One was the simulated experimental system containing microorganisms. Another was the simulated experimental system without microorganisms. Results of previous experiment indicated that the acidic materials and CA like enzymatic materials excreted by fungi and the enwrapping effect of fungal mycelia were important factors influencing limestone dissolution. In the three factors mentioned above, the dissolution effect was mycelia enwraping effect>acidic dissolution effect>CA enzymatic effect. The results of the second experiment demonstrated further that the limestone dissolution effect of the acidic materials excreted by fungi was stronger than that of CA excreted by fungi. Nevertheless, CA still played an important role in promoting the dissolution of limestone. Wei Li and Peng-Peng Zhou have contributed to the article equally.     

Molecular Characterization of an Endolithic Microbial Community in Dolomite Rock in the Central Alps (Switzerland)

Endolithic microorganisms colonize the pores in exposed dolomite rocks in the Piora Valley in the Swiss Alps. They appear as distinct grayish-green bands about 1–8 mm below the rock surface. Based on environmental small subunit ribosomal RNA gene sequences, a diverse community driven by photosynthesis has been found. Cyanobacteria (57 clones), especially the genus Leptolyngbya, form the functional basis for an endolithic community which contains a wide spectrum of so far not characterized species of chemotrophic Bacteria (64 clones) with mainly Actinobacteria, Alpha-Proteobacteria, Bacteroidetes, and Acidobacteria, as well as a cluster within the Chloroflexaceae. Furthermore, a cluster within the Crenarchaeotes (40 clones) has been detected. Although the eukaryotic diversity was outside the scope of the study, an amoeba (39 clones), and several green algae (51 clones) have been observed. We conclude that the bacterial diversity in this endolithic habitat, especially of chemotrophic, nonpigmented organisms, is considerable and that Archaea are present as well.     

Endolithic Phototrophs from an Active Geothermal Region in New Zealand

Endolithic photosynthetic communities in geothermal siliceous rocks in the area of Rotorua, New Zealand, were analysed using traditional microbiological and molecular biology techniques. Rock surface temperatures varied between 40°C and 60°C. Major endoliths included cyanobacteria of subsections I, II and V. Few subsection IV organisms were found and subsection III (filamentous, non-heterocystous) cyanobacteria were present only as epiliths or chasmoendoliths. Therefore, the endolithic cyanobacterial communities in these sites resembled assemblages as reported for carbonate rocks in other geothermal regions. Cells of the rhodophyte family Cyanidiaceae were detected within rock at various sites. Some of these phototrophic organisms were associated with mineral (presumably silica) deposits and could be important geological agents in siliceous rock deposition.     

Fungal Community Analysis by Large-Scale Sequencing of Environmental Samples

Fungi are an important and diverse component of soil communities, but these communities have proven difficult to study in conventional biotic surveys. We evaluated soil fungal diversity at two sites in a temperate forest using direct isolation of small-subunit and internal transcribed spacer (ITS) rRNA genes by PCR and high-throughput sequencing of cloned fragments. We identified 412 sequence types from 863 fungal ITS sequences, as well as 112 ITS sequences from other eukaryotic microorganisms. Equal proportions of Basidiomycota and Ascomycota sequences were present in both the ITS and small-subunit libraries, while members of other fungal phyla were recovered at much lower frequencies. Many sequences closely matched sequences from mycorrhizal, plant-pathogenic, and saprophytic fungi. Compositional differences were observed among samples from different soil depths, with mycorrhizal species predominating deeper in the soil profile and saprophytic species predominating in the litter layer. Richness was consistently lowest in the deepest soil horizon samples. Comparable levels of fungal richness have been observed following traditional specimen-based collecting and culturing surveys, but only after much more extensive sampling. The high rate at which new sequence types were recovered even after sampling 863 fungal ITS sequences and the dominance of fungi in our libraries relative to other eukaryotes suggest that the abundance and diversity of fungi in forest soils may be much higher than previously hypothesized.     

Metagenomic Insights into the Bioaerosols in the Indoor and Outdoor Environments of Childcare Facilities

Airborne microorganisms have significant effects on human health, and children are more vulnerable to pathogens and allergens than adults. However, little is known about the microbial communities in the air of childcare facilities. Here, we analyzed the bacterial and fungal communities in 50 air samples collected from five daycare centers and five elementary schools located in Seoul, Korea using culture-independent high-throughput pyrosequencing. The microbial communities contained a wide variety of taxa not previously identified in child daycare centers and schools. Moreover, the dominant species differed from those reported in previous studies using culture-dependent methods. The well-known fungi detected in previous culture-based studies (Alternaria, Aspergillus, Penicillium, and Cladosporium) represented less than 12% of the total sequence reads. The composition of the fungal and bacterial communities in the indoor air differed greatly with regard to the source of the microorganisms. The bacterial community in the indoor air appeared to contain diverse bacteria associated with both humans and the outside environment. In contrast, the fungal community was largely derived from the surrounding outdoor environment and not from human activity. The profile of the microorganisms in bioaerosols identified in this study provides the fundamental knowledge needed to develop public health policies regarding the monitoring and management of indoor air quality.     

Beringian Paleoecology Inferred from Permafrost-Preserved Fungal DNA

The diversity of fungi in permanently frozen soil from northeastern Siberia was studied by culture-independent PCR amplification of diverse environmental 18S rRNA genes. Elaborate protocols to avoid contamination during drilling, sampling, and amplification were used. A broad diversity of eukaryotic DNA sequences that were 510 bp long, including sequences of various fungi, plants, and invertebrates, could be obtained reproducibly from samples that were up to 300,000 to 400,000 years old. The sequences revealed that ancient fungal communities included a diversity of cold-adapted yeasts, dark-pigmented fungi, plant-parasitic fungi, and lichen mycobionts. DNA traces of tree-associated macrofungi in a modern tundra sample indicated that there was a shift in fungal diversity following the last ice age and supported recent results showing that there was a severe change in the plant composition in northeastern Siberia during this period. Interestingly, DNA sequences with high homology to sequences of coprophilic and keratinophilic fungi indicated that feces, hair, skin, and nails could have been sources of ancient megafauna DNA recently reported to be present in small amounts of Siberian permafrost sediments.     

Beringian paleoecology inferred from permafrost-preserved fungal DNA

The diversity of fungi in permanently frozen soil from northeastern Siberia was studied by culture-independent PCR amplification of diverse environmental 18S rRNA genes. Elaborate protocols to avoid contamination during drilling, sampling, and amplification were used. A broad diversity of eukaryotic DNA sequences that were 510 bp long, including sequences of various fungi, plants, and invertebrates, could be obtained reproducibly from samples that were up to 300,000 to 400,000 years old. The sequences revealed that ancient fungal communities included a diversity of cold-adapted yeasts, dark-pigmented fungi, plant-parasitic fungi, and lichen mycobionts. DNA traces of tree-associated macrofungi in a modern tundra sample indicated that there was a shift in fungal diversity following the last ice age and supported recent results showing that there was a severe change in the plant composition in northeastern Siberia during this period. Interestingly, DNA sequences with high homology to sequences of coprophilic and keratinophilic fungi indicated that feces, hair, skin, and nails could have been sources of ancient megafauna DNA recently reported to be present in small amounts of Siberian permafrost sediments.     

木本植物对喀斯特石质生境岩石形态结构的适应性

为弄清植物对喀斯特岩石形态结构的适应性,选择3种岩石类型及7种木本群落为研究对象,基于分形、空间句法理论研究岩石结构面孔裂隙形态结构及其与群落特征的关系。结果表明:石灰岩结构面孔裂隙间隙度大、白云岩居中、白云质砂岩小,分维数则反之;白云质砂岩结构面孔裂隙整合度、控制值、密度值大,白云岩居中,石灰岩小;白云岩生境下群落物种多样性及优势种碳、氮含量高、白云质砂岩次之、石灰岩低;分形与空间句法指数间呈显著相关关系,两者能较好的揭示岩石形态结构;总体上3种岩性生境下物种多样性及优势种碳、氮含量与岩石结构面孔裂隙分形及空间句法指数具较强相关关系;大致相同的环境下白云岩形态结构适宜植物生长、白云质砂岩次之、石灰岩差;研究岩石形态结构对喀斯特植被恢复具有重要意义。     

不同风化年限碳酸岩风化壳真菌群落结构特征

【目的】岩生真菌是促进碳酸岩生物风化的重要推动者,研究黔中典型喀斯特区不同风化年限碳酸岩风化壳真菌群落结构特征有利于了解真菌对岩石的风化作用。【方法】选取位于黔中花溪区南部的废弃碳酸岩墓碑为调查对象,对不同风化年限碳酸岩风化壳进行采样,运用宏基因组测序方法对风化壳样品进行基因测序,并利用统计学方法对真菌群落结构特征及功能特征进行分析。【结果】18个风化壳样品中共获得1087种真菌,分属于9个门、44纲、538属。不同样本之间真菌群落数量和组成差异较大,在碳酸岩风化过程中,子囊菌门(Ascomycota)始终为优势类群,平均相对丰度达到95%以上,随风化年限的增加呈现显著下降的趋势。Shannon指数和Simpson指数显示,碳酸岩风化壳真菌群落多样性随风化年限的增加呈现先减小后增加再减小的趋势。从所有样本中共检测出3 379 478个KEGG pathway level 3通路相关基因,主要与物质能量的代谢、运输等功能相关。与碳循环、氮循环和硫循环相关的主要微生物属于子囊菌门,随着风化年限的增加呈现下降的趋势。冗余分析(redundancyanalysis,RDA)结果表明,三氧化二铁...     

Endolithic fungi in marine ecosystems

Fungi are an important constituent of microbial endolithic assemblages in marine ecosystems. As euendoliths, they penetrate limestone, mollusk shells and other carbonate substrates, where they can exploit mineralized organic matter, attack their hosts, or engage in symbiotic relationships. They leave specific boring traces, which can be identified in the fossil record and described as trace fossils. Their distribution is independent of light and extends from the intertidal ranges to abyssal oceanic depths. Important, but insufficiently studied, is the role of aggressive endolithic fungi in skeletons of corals where they are ubiquitous and globally distributed. In healthy growing reef corals, the relationship between the coral coelenterate, endolithic algae and fungi is in a state of equilibrium, but can turn detrimental to coral health when reefs are exposed to environmental stress.     

贡嘎蝠蛾幼虫肠道真菌多样性分析

[目的]分析贡嘎蝠蛾肠道(Hepialus gonggaensis)幼虫肠道真菌多样性.[方法]采用常规分离与分子鉴定的方法和基于ITS(internal transcribed spacer)基因的RFLP方法,建立贡嘎蝠蛾幼虫肠道真菌的ITS克隆文库,分别用MspⅠ、HaeⅢ和Taq Ⅰ对205个阳性克隆的质粒酶切指纹图谱分析,结果显示有23个不同的RFLP操作分类单元(OTU),对这23个操作分类单元的阳性克隆子进行测序并绘制系统进化树.[结果]结果显示贡嘎蝠蛾幼虫肠道内存在8个属的真菌类群.其中被孢霉属(Mortierella)和丝孢酵母属(Trichosporon)的丰度最高,分别占克隆文库的46.34%和40.00%,鉴定为肠道内的优势真菌类群.用常规分离与分子鉴定方法只获得隐球酵母(Cryptococcus magnus)、Geomyces sp和丝孢酵母(Trichosporon porosum)3个类群的真菌.结合常规分离法与RFLP法能够更有效的分析肠道微生物的多样性,获得更多更全面的微生物多样性信息.